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Preparation method and application of conditioned medium for deer periosteum

A culture medium and deer bone technology, applied in vertebrate cells, artificial cell constructs, animal cells, etc., can solve the problems of inability to provide a culture environment for tissues, inability to collect conditioned medium at any time, and difficulty in controlling the concentration of carbon dioxide. The effect of culture time, improving survival rate and convenient operation

Inactive Publication Date: 2014-11-19
INST OF SPECIAL ANIMAL & PLANT SCI OF CAAS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This kind of culture cannot provide a three-dimensional culture environment for the tissue, and it is difficult to control the carbon dioxide concentration during the culture process, and the conditioned medium cannot be collected at any time during the culture process

Method used

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  • Preparation method and application of conditioned medium for deer periosteum
  • Preparation method and application of conditioned medium for deer periosteum
  • Preparation method and application of conditioned medium for deer periosteum

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] The invention provides a method for preparing conditioned medium by cultivating staghorn periosteum tissue in a triangular flask, the steps are as follows:

[0041] 1) Using animal tissue cutter ( figure 1 ) cutting the deer periosteum tissue collected from the living body into thin slices with a length of 6 mm, a width of 2 mm, and a thickness of 1.7 mm to obtain slices of the periosteum tissue;

[0042] 2) Slice 20 pieces of periosteum tissue obtained in step 1), and put them into a Erlenmeyer flask.

[0043] 3) Add nerve culture medium to the Erlenmeyer flask and fill it with saturated concentration of CO 2 Concentration, stopper the Erlenmeyer flask, put it in the incubator and cultivate it for 4 days, and collect the culture medium after the cultivation is completed;

[0044] 4) Filter the culture solution obtained in step 3) with a 0.22 μm filter membrane to obtain the velvet antler periosteum tissue-conditioned culture solution.

Embodiment 2

[0046] The invention provides a method for preparing the conditioned culture medium of periosteum tissue in deer antler area, the steps are as follows:

[0047] 1) Using an animal tissue cutter, cut the periosteum tissue from the deer antler area collected from the living body into thin slices with a length of 3 mm, a width of 2 mm, and a thickness of 0.7 mm to obtain slices of the periosteum tissue;

[0048] 2) Put 15 pieces of periosteum tissue slices obtained in step 1) evenly into an insert dish coated with rat tail collagen, and then put the insert dish into a 24-well culture plate ( figure 2 a) in the hole;

[0049] 3) Add nerve culture medium to the insert dish and the 24-well culture plate, that is, the upper chamber and the lower chamber of the culture, and store at 37°C, CO 2 Cultivate for 3 days under the condition of 5% concentration and saturated humidity, and collect the culture medium after the cultivation is completed;

[0050] 4) Filter the culture solution...

Embodiment 3

[0052] The invention provides a method for preparing conditioned culture medium of staghorn periosteum tissue, the steps are as follows:

[0053] 1) Use an animal tissue cutter to cut the periosteum tissue collected from the living body into thin slices with a length of 3 mm, a width of 2 mm, and a thickness of 0.7 mm to obtain periosteum tissue slices;

[0054] 2) Slice 50 pieces of periosteal tissue obtained in step 1), put them into an insert dish coated with rat tail collagen, and put the insert dish into the well of a 6-well culture plate;

[0055] 3) To the insert dish and 6-well culture plate ( figure 2 b) Add nerve culture medium, at 37°C, CO 2 Cultivate for 3 days under the condition of 5% concentration and saturated humidity, and collect the culture medium after the cultivation is completed;

[0056] 4) Filter the culture solution obtained in step 3) with a 0.22 μm filter membrane to obtain the velvet antler periosteum tissue-conditioned culture solution.

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Abstract

The invention discloses a preparation method and application conditioned medium for deer periosteum, and belongs to technical field of cell tissue culture. The method provided by the invention is as below: inserting deer periosteum tissue slice in an insert dish, adding medium into the insert dish, collecting culture solution after culture, filtering to obtain conditioned medium. The insert dish is composed of an upper chamber and a lower chamber, and is placed into a porous culture plate. The method provided by the invention can make the deer periosteum tissue secrete active substances under in vitro culture conditions, so as to collect and analyze the secreted active substances. The method solves the problems that in vitro culture cannot provide three-dimensional cultivation environment for the tissue, carbon dioxide concentration is difficult to control in the cultivation process of and conditioned medium can not be collected momentarily. At the same time, the method provided by the invention effectively improves the survival rate of cultured tissue, shortens the culture time of the tissue, and can collect conditioned medium at any time.

Description

technical field [0001] The invention relates to a preparation method and application of deer periosteum conditioned culture solution, belonging to the technical field of cell tissue culture. Background technique [0002] Antler is the only complex mammalian organ known to be capable of complete regeneration of cartilage, bone, skin, blood vessels, and nerves. Among them, the research on nerve regeneration found that the regeneration and rapid growth of pilose antler nerves are mainly achieved by the stimulation of biomolecules derived from the periosteum of the horn handle. However, in order to analyze the biomolecules derived from the horn-stalk periosteum, an in vitro culture system must be established. [0003] In the process of cell culture, cells may secrete active substances into the culture medium. After culturing certain cells or tissues, the culture medium containing secretions from cells or tissues is called conditioned medium. At present, there are many studies ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071
Inventor 李春义王文英孙红梅
Owner INST OF SPECIAL ANIMAL & PLANT SCI OF CAAS
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